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Prnp knockdown in transgenic mice using RNA interference

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Abstract

RNA interference has become a widely used approach to perform gene knockdown experiments in cell cultures and more recently transgenic animals. A designed miRNA targeting the prion protein mRNA was built and expressed using the human PRNP promoter. Its efficiency was confirmed in transfected cells and it was used to generate several transgenic mouse lines. Although expressed at low levels, it was found to downregulate the endogenous mouse Prnp gene expression to an extent that appears to be directly related with the transgene expression level and that could reach up to 80% inhibition. This result highlights the potential and limitations of the RNA interference approach when applied to disease resistance.

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Abbreviations

dsRNA1:

Double-stranded RNA

miRNA:

Micro RNA

PrPC :

Cellular PrP protein

pre-miRNA:

Precursor of miRNA

RNAi:

RNA interference

siRNA:

Small interfering RNA

shRNA:

Short hairpin RNA

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Acknowledgements

We are most grateful to Dr. J. Collinge (National Hospital for Neurology and Neurosurgery, London, UK) for the kind gift of the pMG5 plasmid, to Prof. Bujard (ZMBH, Heidelberg, Germany) for that of pUHD 10.3. Micaela Gallozzi is a Ph.D student supported by the European Community RIVAGE Marie Curie Action. This work was partially supported by the ANR-05, GenAnimal VecteurshRNA.

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Authors and Affiliations

  1. UR 339, Laboratoire de Génétique Biochimique et de Cytogénétique, Institut National de la Recherche Agronomique, 78350, Jouy-en-Josas, France

    Micaela Gallozzi, Fabienne Le Provost, Caroline Morgenthaler, Marthe Vilotte, Bruno Passet, José Costa, Gaëlle Tilly & Jean-Luc Vilotte

  2. UR 892, Unité de Virologie et Immunologie Moléculaires, Institut National de la Recherche Agronomique, 78350, Jouy-en-Josas, France

    Jérome Chapuis, Annick Le Dur, Laetitia Herzog, Vincent Beringue, Philippe Tixador & Hubert Laude

  3. UE 907, Unité Expérimentale Rongeurs, Institut National de la Recherche Agronomique, 78350, Jouy-en-Josas, France

    Coralie Peyre

  4. UMR 1198, Unité de Biologie du Développement et de la Reproduction, Institut National de la Recherche Agronomique, 78350, Jouy-en-Josas, France

    Nathalie Daniel-Carlier & Eric Pailhoux

Authors
  1. Micaela Gallozzi
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  2. Jérome Chapuis
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  3. Fabienne Le Provost
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  4. Annick Le Dur
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  5. Caroline Morgenthaler
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  7. Nathalie Daniel-Carlier
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  8. Eric Pailhoux
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  9. Marthe Vilotte
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  10. Bruno Passet
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  11. Laetitia Herzog
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  14. Philippe Tixador
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  15. Gaëlle Tilly
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  16. Hubert Laude
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  17. Jean-Luc Vilotte
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Correspondence to Jean-Luc Vilotte.

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Gallozzi, M., Chapuis, J., Le Provost, F. et al. Prnp knockdown in transgenic mice using RNA interference. Transgenic Res 17, 783–791 (2008). https://doi.org/10.1007/s11248-008-9179-2

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